Journal
MOLECULAR CELL
Volume 81, Issue 10, Pages 2216-+Publisher
CELL PRESS
DOI: 10.1016/j.molcel.2021.03.032
Keywords
-
Categories
Funding
- ZonMW TOP grant [91215067]
- European Research Council (ERC) Advanced Grant [694466]
- NIH Common Fund 4D Nucleome'' Program grant [U54DK107965]
- Marie Curie/AIRC iCARE2.0 fellowship [800924]
- KWF Dutch Cancer Society
Ask authors/readers for more resources
Through systematically analyzing the impact of chromatin context on three DNA double-strand break repair pathways, it was found that non-homologous end-joining is more common in euchromatin, while microhomology-mediated end-joining is more active in specific heterochromatin contexts. Additionally, single-stranded template repair competes strongly with other repair pathways and is moderately linked to chromatin context.
DNA double-strand break (DSB) repair is mediated by multiple pathways. It is thought that the local chromatin context affects the pathway choice, but the underlying principles are poorly understood. Using a multiplexed reporter assay in combination with Cas9 cutting, we systematically measure the relative activities of three DSB repair pathways as a function of chromatin context in >1,000 genomic locations. This reveals that non-homologous end-joining (NHEJ) is broadly biased toward euchromatin, while the contribution of microhomology-mediated end-joining (MMEJ) is higher in specific heterochromatin contexts. In H3K27me3-marked heterochromatin, inhibition of the H3K27 methyltransferase EZH2 reverts the balance toward NHEJ. Single-stranded template repair (SSTR), often used for precise CRISPR editing, competes with MMEJ and is moderately linked to chromatin context. These results provide insight into the impact of chromatin on DSB repair pathway balance and guidance for the design of Cas9-mediated genome editing experiments.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available